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MC38-N4/OT-I co-culture system screening method and application of screened polysaccharide composition and polysaccharide composition

A technology of MC38-N4, 1.MC38-N4, applied in the field of functional food, can solve the problems of invisibility of experimental results, radioactive toxicity, cumbersome experimental process, etc., to improve the immune function of the body, promote the killing effect, and have a wide range of applications. Effect

Pending Publication Date: 2022-04-15
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

in, 51 The Cr release assay is the most common in vitro assay and is commonly used to measure T cell-mediated cytotoxicity, but its application is limited due to its radiotoxicity
Pimentel et al. proposed to use the firefly luciferase assay to evaluate the targeted killing mortality of tumor cells. The need for experimental large-scale screening (Olivo PimentelV, Yaromina A, Marcus D, Dubois LJ, Lambin P.A novel co-culture assay to assessanti-tumor CD8+T cell cytotoxicity via luminescence and multicolor flowcytometry.J Immunol Methods.2020Dec;487:112899 .doi:10.1016 / j.jim.2020.112899.Epub 2020Oct 15.PMID:33068606.)

Method used

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  • MC38-N4/OT-I co-culture system screening method and application of screened polysaccharide composition and polysaccharide composition
  • MC38-N4/OT-I co-culture system screening method and application of screened polysaccharide composition and polysaccharide composition
  • MC38-N4/OT-I co-culture system screening method and application of screened polysaccharide composition and polysaccharide composition

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Experimental program
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preparation example Construction

[0038] The preparation methods of different raw material extracts include:

[0039] 1. The preparation method of Agaricus blazei polysaccharide extract

[0040] Dry Agaricus blazei to constant weight, and grind the dry raw materials into powder, and pass through a 200-mesh sieve. Add distilled water according to the ratio of solid to liquid 1:20 (m / m), extract at 55°C for 2 hours, and the number of extractions is 2 times, combine the two extracts, centrifuge the extract at 4000r / min for 10min, and collect the supernatant The solution was concentrated by rotary evaporation at 55°C under reduced pressure. Add 3 times the volume of the concentrated solution of absolute ethanol, let stand at 4°C for 24 hours, centrifuge at 4000r / min for 15 minutes, collect the precipitate, and vacuum freeze-dry to obtain Agaricus blazei crude polysaccharide.

[0041] 2, the preparation method of Grifola frondosa polysaccharide extract

[0042] Grifola frondosa is dried to constant weight, the d...

Embodiment 1

[0051] The polysaccharide composition was prepared according to the following ratio: the mass-number ratio of Agaricus blazei polysaccharide, Grifola frondosa polysaccharide, Tangerine peel polysaccharide and Phellinus polysaccharide was 125 parts: 1 part: 1 part: 0 parts.

Embodiment 2

[0052] Embodiment 2 (comparative example 6)

[0053] The polysaccharide composition is prepared according to the following ratio: the mass-number ratio of Agaricus blazei polysaccharide, Grifola frondosa polysaccharide, Phellinus polysaccharide and Tangerine peel polysaccharide is 1 part: 1 part: 5 parts: 1 part.

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Abstract

The invention discloses a screening method of an MC38-N4 / OT-I co-culture system and application of a screened polysaccharide composition and a polysaccharide composition. The polysaccharide composition obtained based on the screening method comprises the following steps: obtaining tumor target cells containing specific antigens and fluorescent proteins, separating immune effector cells, co-culturing the tumor target cells and the immune effector cells, and evaluating the immunomodulatory function of the polysaccharide composition. The co-culture screening method provided by the invention has the advantages that the interaction process of tumor cells and immune cells is simulated based on the characteristics of a tumor microenvironment, and the co-culture screening method has the characteristics of wide application range, visual experimental results, high-throughput screening and the like. The polysaccharide composition provided by the invention is prepared from the following raw materials: agaricus blazei murill, grifola frondosa and pericarpium citri reticulatae, has the effects of improving the body immune function, enhancing the anti-tumor effect and the like, can be used for developing immune enhancement related products, and provides an experimental basis and a scientific basis for applying a food-borne polysaccharide composition to clinical immune enhancement medicines.

Description

technical field [0001] The invention belongs to the technical field of functional food, and specifically relates to a screening method for MC38-N4 / OT-I co-cultivation system and the application of the screened polysaccharide prescription and polysaccharide composition. Background technique [0002] The occurrence, growth and metastasis of tumors are closely related to the internal and external environment of tumor cells. Tumor micro-environment (TME) refers to the surrounding microenvironment where tumor cells exist, including surrounding blood vessels, immune cells, fibroblasts, bone marrow-derived cells, various signaling molecules and extracellular matrix. In recent years, because immunotherapy can effectively stimulate the host immune system and promote immune cells in the tumor microenvironment to kill cancer cells, it has developed into the fourth anti-tumor treatment after surgery, chemotherapy, and radiotherapy in cancer treatment. Studies have found that natural po...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078C12N5/09C12Q1/02A61K31/715A61P35/00A61P37/04
Inventor 任娇艳徐天雄姚茂金黄楚君
Owner SOUTH CHINA UNIV OF TECH